We will study the effect of diethylstilbestrol (DES) administration to immature chicks on the assembly of chick oviduct lipid intermediates in glycoprotein glycosylation. In particular the assembly of the oligosaccharide lipid will be studied by incubation of oviduct membrane preparations with UDP-(14C)GlcNAc, pulse-chase experiments with UDP-(14C)-GlcNAc and unlabeled GDP-Man, as well as incubation of isolated radioactive intermediates in substrate level quantities with membranes. We will also determine the effect of DES treatment on the levels of endogenous dolichylphosphate in the oviduct preparations by partial purification of dolichylphosphate from oviducts and measuring by an enzymatic assay. If dolichylphosphate levels in the preparation change in response to DES treatment, we will attempt to determine the mechanism of that change. Other hormonal manipulations, such as withdrawal from DES, withdrawal followed by secondary stimulation, and progesterone adminstration will be performed. During these treatments we will measure dolichylphosphate levels and attempt to determine the mechanism of these changes. The proteins glycosylated by oviduct preparation using GDP-Man as substrate appear to change during the course of DES treatment. We will study those changes in more detail using two-dimensional electrophoresis and double-label experiments with (3H) and (14C)GDP-Man. In addition to changes in the lipid intermediate pathway upon various hormonal treatments, we have observed changes in glycosidases, particularly beta-N-acetylglucosaminidase. The N-acetylglucosamindase is found in at least two isozymic forms that may be differentially regulated by hormonal treatment. The oviduct cells responsible for the synthesis of each of these isozymes will be localized using fluorescent antibodies against the purified isozymes. We will also try to determine the function of each of the isozymes.